Figure 2.

MPP+-induced cell death is not dependent on necroptosis. (a and b) After treatment of SH-SY5Y cells with RA, RIP1 short hairpin RNA (shRNA) was induced by incubation with Dox (2 μg/ml) for 5 days in the presence of BDNF. (a) Dox-treated neuronal cells were incubated with MPP+ (5 mM) for 48 h in the presence or absence of Nec-1 (20 μM). Cell death was calculated from lactate dehydrogenase (LDH) leakage. (b) RIP1 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were detected by western blotting. The extreme right lane is Jurkat cell lysate (positive control). (c) Cells were treated with MPP+ in the presence of Nec-1 or its derivatives (20 μM each) for 48 h. Cell death was calculated from LDH leakage. (d and e) After treatment of SH-SY5Y cells with RA, RIP3 shRNA was induced by incubation with Dox (2 μg/ml) for 5 days in the presence of BDNF. (d) Dox-treated neuronal cells were incubated with MPP+ (5 mM) for 48 h. Cell death was calculated from LDH leakage. (e) RIP3 and GAPDH were detected by western blotting. The extreme right lane is Jurkat cell lysate (positive control). Data are shown as the mean±S.D. of three independent experiments. NS, not significant; **P<0.01.